, Volume 256, Issue 1, pp 84-87

Single-step conjugative cloning of bacterial gene fusions involved in microbe-host interactions

Rent the article at a discount

Rent now

* Final gross prices may vary according to local VAT.

Get Access

Abstract

In vivo expression technology (IVET) is a genetic strategy for isolating genes expressed in vivo. In order to fully exploit this technology, it is necessary to analyse large numbers of IVET-generated gene fusions, which must be recovered from the chromosome of host bacteria. In bacteria for which transductional methods are not available, the recovery of integrated fusion plasmids is problematic and currently limits broad application of IVET. We describe a rapid, single-step, triparental conjugative approach for recovering chromosomally integrated fusion plasmids from both Pseudomonas fluorescens and Salmonella typhimurium. This simple and broadly applicable conjugative cloning system extends the utility of the IVET approach to clinically and agronomically relevant microbes and may be employed to recover non-replicating and integrated plasmids in other systems.

Received: 28 March 1997 / Accepted: 23 May 1997